D-β-lysylmethanediamine derivatives and preparation thereof

ABSTRACT

Now are provided (R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamides which are novel compounds having the general formula (I): ##STR1## wherein n stands for 2-5, and which may be for example, (R)-3,6-diamino-N-(2-aminoethyl)hexanamide (n=2) and (R)-3,6-diamino-N-(3-aminopropyl)hexanamide (n=3). Their preparation process is also provided. These novel compounds and acid addition salts thereof have activities inhibitory against Gram-positive bacteria, Gram-negative bacteria and AIDS virus, as well as tumor cells and are chemically stable. 
     These novel compounds and their salts are useful as chemotherapeutic agents for diseases caused by these bacteria or virus and also as antitumor agent.

This application is a 371 of PCT/JP95/01680, filed Aug. 24, 1995.

TECHNICAL FIELD

This invention relates to an (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamidewhich is a novel compound active for inhibiting the growth ofGram-positive bacteria and Gram-negative bacteria, the infection ofhuman T-cells with AIDS virus, namely HIV, and the growth of tumor cellsand which is represented by the following general formula (I): ##STR2##wherein n stands for an integer of 2-5. This novel compound of thegeneral formula (I) above is useful as a chemotherapeutic agent forvarious purposes. This invention also relates to a process for thepreparation of the above-described novel compound.

BACKGROUND ART

The present inventors discovered that D-β-lysyl-methanediamine whichcorresponds to such compound of the general formula (I) above where nstands for 1 and is hence represented by the formula (A): ##STR3## isproduced and accumulated in a culture of an actino-mycetes strain ("J.Antibiotics", 39, 476(1986) and Japanese Patent Application Laid-OpenPublication No. SHO 62-114947, published May 26, 1987). Development andresearches are now made for utilization of D-β-lysyl-methanediamine as atherapeutic agent for the treatment of AIDS (Japanese Patent ApplicationNo. HEI 3-346341 filed on Dec. 27, 1991; U.S. patent application Ser.No. 08/256,121; and European patent application publication No.653207-A1).

AIDS is a disease caused by the infection with an AIDS virus (HIV).Several drugs which can inactivate HIV have been reported astherapeutics for AIDS, but any of them are not satisfactory. There ishence a continued demand for providing such a novel drug having lowtoxicity but high anti-HIV activities.

Through a study of the present inventors, the chemical structure ofD-β-lysylmethanediamine (which is generally called "Bellenamine") wasdetermined to be unique since its total synthesis was somewhat difficultwith starting from an amino-protected D-β-lysylglycine via its acidazide "J. Antibiotics", 45, 1677(1992)!. Owing to the high instabilityof D-β-lysylmethanediamine, its total synthesis process is troublesometo be achieved.

D-β-lysylmethanediamine, that is, Bellenamine, which was found in theculture of the microorganism of actino-mycetes and discovered by thepresent inventors and which has anti-HIV activities, has the linearaldoaminal structure that is discovered firstly to exist in the nature,so that Bellenamine is chemically instable.

DISCLOSURE OF THE INVENTION

The present inventors therefore have now conducted research on thesynthesis of novel and useful Bellenamine-related substances. An objectof the present invention is to provide an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide as a novel compound which canbe considered to be analogous to D-β-lysylmethanediamine but ischemically more stable. Another objects of the present invention will beclear from the following descriptions.

The present inventors have now succeeded in synthesizing a series of(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamides, which each is a novelcompound having higher chemical stability than D-β-lysylmethanediamineand having antibacterial activity and anti-HIV activities as well asantitumor activity and which is collectively represented by the generalformula (I) as above. It has now been found that among the compounds ofthe general formula (I), (R)-3,6-diamino-N-(2-aminoethyl)hexanamide and(R)-3,6-diamino-N-(3-aminopropyl)hexanamide have antibacterial andanti-HIV activities comparable to or sometime superior to those ofD-β-lysylmethanediamine and also exhibit an antitumor activity superiorto that of D-β-lysylmethanediamine.

According to a first aspect of the present invention, therefore, thereis provided an (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide represented bythe general formula (I): ##STR4## wherein n stands for an integer of2-5, or an acid addition salt thereof.

According to a second aspect of the present invention, there is provideda process for the preparation of an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide represented by the generalformula (I): ##STR5## wherein n stands for an integer of 2-5, whichcomprises the steps of condensing, through an amido-bond, amono-amino-protected derivative of an α,ω-alkanediamine represented bythe general formula (II):

    H.sub.2 N(CH.sub.2).sub.n NH-A                             (II)

wherein A represents an aralkyloxycarbonyl group which is anamino-protecting group easily removable by hydrogenolysis and n standsfor an integer of 2-5, with a bis(N-aralkyloxycarbonyl)-D-β-lysinerepresented by the general formula (III): ##STR6## wherein A has thesame meaning as described above, to form an(R)-3,6-bis(aralkyloxycarbonylamino)-N-(ω-aralkyloxycarbonylaminoalkyl)hexanamiderepresented by the general formula (IV): ##STR7## wherein A and n havethe same meanings as described above, and then subjecting the compoundof the formula (IV) to hydrogenolysis to remove the aralkyloxycarbonylgroups therefrom.

The (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide of the general formula(I) synthesized as a novel compound according to the present inventionor the acid addition salt thereof are chemically stable, and besidesthey have inhibitory activities against Gram-positive bacteria andGram-negative bacteria and also against AIDS virus, as well as anantitumor activity against some kinds of tumor cells, so that they areuseful as a chemotherapeutic agent for diseases caused by infection withthese bacteria or virus and also are useful as an antitumor agent.

BEST MODE FOR CARRYING OUT THE INVENTION

A detailed description will hereinafter be made of the first aspect ofthe present invention.

When n stands for 2 in the compound of the general formula (I), there isprovided (R)-3,6-diamino-N-(2-aminoethyl)hexanamide represented by thefollowing formula (Ia): ##STR8##

When n stands for 3 in the compound of the general formula (I), there isprovided (R)-3,6-diamino-N-(3-aminopropyl)hexanamide represented by thefollowing formula (Ib): ##STR9##

Physicochemical properties of some examples of the novel compounds ofthe general formula (I) available according to the present inventionwill be described below.

1! (R)-3,6-Diamino-N-(2-aminoethyl)hexanamide the compound of theformula (Ia)!

(1) Color and appearance: a colorless paste

(2) Molecular formula: C₈ H₂₀ N₄ 0

(3) Mass spectrum (FD-MS): m/z 189 (MH⁺ High-resolution mass spectrum(HR-FAB-MS): m/z 189.1729 (MH⁺), Calculated for C₈ H₂₁ N₄ 0: 189.1715(MH)

(4) Melting point: No definite melting point is observed.

(5) Specific optical rotation: α!_(D) ²⁴ -6.0° (c 1.2, H₂ O)

(6) Infrared absorption spectrum (KBr): 3300, 2950, 1640, 1570, 1480,1440, 1390, 1320, 1240, 1160, 1050, 820 cm⁻¹

(7) ¹ H-NMR spectrum (400 MHz, D₂ O, pD4):

δ1.80(4 H, m, 4-H₂, 5-H₂)

2.67(1 H, dd, J=16.4, 8.5 Hz, 2-H)

2.80(1 H, dd, J=16.4, 4.6 Hz, 2-H)

3.07(2 H, m, 6-H₂)

3.18(2 H, t, J=5.9 Hz, 2'-H₂)

3.55(2 H, t, J=5.9 Hz, 1'-H₂)

3.71(1 H, m, 3-H)

(8) ¹³ C-NMR spectrum (100 MHz, D₂ O, pD 4):

δ23.8(t, C-5), 30.0(t, C-4), 37.3(t, C-2),

37.7(t, C-1'), 39.8(t, C-6), 40.0(t, C-2"),

49.1(d, C-3), 173.6(s, C-1)

(9) Solubility: highly soluble in water.

(10) Basic, acidic or neutral: basic substance.

2! (R)-3,6-Diamino-N-(3-aminopropyl)hexanamide the compound of theformula (Ib)!

(1) Color and appearance: a colorless paste

(2) Molecular formula: C₉ H₂₂ N₄ O

(3) Mass spectrum (FD-MS): m/z 203 (MH⁺

High-resolution mass spectrum (HR-FAB-MS):

m/z 203.1880 (MH⁺),

Calculated for C₉ H₂₃ N₄ O: 203.1872 (MH)

(4) Melting point: No definite melting point is observed.

(5) Specific optical rotation: α!_(D) ²⁴ -3.5°(c 0.9, H₂ O)

(6) Infrared absorption spectrum (KBr): 3300, 2950, 1640, 1560, 1480,1440, 1390, 1320, 1210, 1150, 1050, 820 cm¹

(7) ¹ H-NMR spectrum (400 MHz, D₂ O, pD4):

δ1.79(4 H, m, 4-H₂, 5-H₂)

1.92(2 H, tt, J=7.3, 7.3Hz, 2'-H₂)

2.65(1 H, dd, J=16.4, 8.1Hz, 2-H)

2.76(1 H, dd, J=16.4, 5.1Hz, 2-H)

3.05(2H, t, J=7.3Hz, 3'-H₂)

3.07(2H, m, 6-H₂)

3.33(2H, m, 1'-H₂)

3.68(1 H, m, 3-H)

(8) ¹³ C-NMR spectrum (100 MHz, D₂ O, pD 4):

δ23.7(t, C-5), 27.4(t, C-2'), 29.9(t, C-4),

37.0(t, C-1'), 37.4(t, C-2), 37.9(t, C-3'),

39.7(t, C-6), 49.2(d, C-3), 172.8(s, C-1)

(9) Solubility: highly soluble in water.

(10) Basic, acidic or neutral: basic substance.

Each compound of the general formula (I) according to the presentinvention is strongly basic so that it can form an addition salt with anacid. Illustrative acids for the formation of acid addition salts mayinclude pharmaceutically acceptable inorganic acids such as hydrochloricacid, sulfuric acid and carbonic acid; and pharmaceutically acceptableorganic acids such as acetic acid, malic acid, citric acid, ascorbicacid and methanesulfonic acid.

Each compound of the general formula (I) according to the presentinvention or an acid addition salt thereof can be administered in theform of a preparation as mixed with a pharmaceutically acceptable solidor liquid carrier, and for example, may be given orally in the dosageform of powder, granules, tablets, syrup or the like, or parenterally inthe dosage form of an injection or the like.

The biological properties of the novel(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide available according to thepresent invention are described below.

1! Antibacterial activities

The minimum inhibitory concentrations (MIC.) of(R)-3,6-diamino-N-(2-aminoethyl)hexanamide of the formula (Ia) and(R)-3,6-diamino-N-(3-aminopropyl)hexanamide of the formula (Ib) againstthe growth of various bacteria were determined by a serial dilutionmethod on a 0.5% peptone agar medium (incubated at 37° C. for 18 hours).The results obtained are shown in Table 1 below, in comparison with theMIC. of D-β-lysylmethanediamine (Bellenamine).

                  TABLE 1                                                         ______________________________________                                                       MIC (μg/ml)                                                                  Bellenamine                                                                             Invention                                                                              Invention                                 Test             (Com-     compound compound                                  Bacteria         parative) (Ia)     (Ib)                                      ______________________________________                                        Staphylococcus aureus FDA209P                                                                  50        25       200                                       S. aureus Smith  6.25      6.25     50                                        Micrococcus luteus FDA16                                                                       >200      >200     >200                                      Bacillus anthracis                                                                             100       100      200                                       B. subtilis NRRL B-558                                                                         12.5      25       100                                       B. subtilis PC1219                                                                             12.5      25       100                                       B. cereus ATCC10702                                                                            50        50       200                                       Escherichia coli NIHJ                                                                          200       100      200                                       E. coli K-12     >200      200      200                                       Shigella dysenteriae JS11910                                                                   >200      200      200                                       S. flexnelli 4b JS1811                                                                         >200      100      200                                       Klebsiella pneumoniae PCI602                                                                   >200      200      >200                                      Proteus vulgaris OX19                                                                          >200      200      200                                       P. mirabilis IFM OM-9                                                                          200       25       50                                        Providencia rettgeri GN311                                                                     50        25       100                                       P. rettgeri GN466                                                                              100       50       50                                        Serratia marcescens                                                                            200       100      200                                       Pseudomonas aeruginosa A3                                                                      200       100      200                                       ______________________________________                                    

2! Inhibitory activities against HIV infection In the following, thereis described Test 1 which demonstrates the inhibitory activities of(R)-3,6-diamino-N-(2-aminoethyl)hexanamide (Ia) and(R)-3,6-diamino-N-(3-aminopropyl)hexanamide (Ib) against the infectionof human T-cells with HIV, namely the AIDS virus.

Test 1

To a Costar 48-well plate, there were added 0.5 ml of a cell suspensionof MT-4 cells, that is, a kind of the human T-cells (containing 1×10⁵cells/ml of a phosphate buffer) and 0.05 ml of a solution (in aphosphate buffer) containing a given amount of a tested compound per onewell. After incubation for 2 hours at 37° C. in an incubator under 5%CO₂, the MT-4 cells were infected with 0.05 ml of a suspension of HIV(HTLV-III_(B) strain) in an amount of multiplicity of infection (m.o.i)of 0.025-0.05, followed by further incubation for 4 days.

Portions of the incubated cultures were taken and the MT-4 cells weresmeared onto slide glasses, dried and immobilized with acetone. Thepresence of the HIV antigen-positive MT-4 cells was detected by theindirect immunofluoroescent antibody assay method Y. Hinuma et al.,"Proc. Natl. Acad. Sci. USA," 78, 6476-6480, (1981) and Y. Takeuchi etal., "Gann.", 78, 11-15 (1987)!. To this end, the cell smears, that is,the acetone-immobilized cell smears were treated at 37° C. for 30minutes with such serum of AIDS patient at a dilution of 1:10 inphosphate buffered saline, which was employed as the first antibody.After washing subsequently with phosphate buffered saline, the MT-4cells were treated at 37° C. for 30 minutes with fluorescentisothiocyanate-conjugated rabbit anti-human IgG serum (CappelLaboratories, Cocharanville, Pa., USA), which was employed as the secondantibody. After the cell smears were then washed with phosphate bufferedsaline and covered with a cover glass, the MT-4 cells were examinedunder a fluorescence microscope. Percentages of the number of the viralantigen-positive MT-4 cells (namely, immunofluorescent cells where theHIV-associated antigens were present and expressed) in total cells werecalculated.

For the control tests of estimating the number of the total cells undertest, the above test procedures were repeated without addition of thetested compound.

Furthermore, cytotoxicity of the tested compound to the MT-4 cells wasestimated. This was done by incubating the MT-4 cells at varyingconcentrations of the tested compound and in the absence of HIV but inthe same manner of incubation and under the same conditions ofincubation of MT-4 cells as those employed in the above-mentioned testprocedures of assaying the activity of the tested compound to inhibitthe infection of T-cells with HIV. That the cytotoxicity was notobserved is represented by the symbol "-" in Table 2 below.

In order to evaluate the activity of the tested compound (Ia) or (Ib) toinhibit the infection of the MT-4 cells with HIV, calculation was madeof the percentages (T/C, %) of the number (T) of the HIVantigen-positive cells as measured in the above-mentioned tests wherethe incubation of MT-4 cells was effected in the presence of the testedcompound, against the number (C) of the HIV antigen-positive cells asmeasured in the above-mentioned control tests where the incubation ofMT-4 cells was effected without the addition of the tested compound. Theresults of such calculation of the T/C values (%) are shown in Table 2below, in term of rate (%) of presence or occurrence of the HIVantigen-positive cells.

                  TABLE 2                                                         ______________________________________                                                  Rate (%) of presence of HIV                                                   antigen-positive cells, and                                                   Cytotoxicity (given in brackets)                                    Concentration of         Invention                                                                              Invention                                   test compound                                                                             Bellenamine  compound compound                                    (μg/ml)  (Comparative)                                                                              (Ia)     (Ib)                                        ______________________________________                                        0           90           90       90                                                      (-)          (-)      (-)                                         0.1         50           90       80                                                      (-)          (-)      (-)                                         1           5            80       10                                                      (-)          (-)      (-)                                         10          10           30        5                                                      (-)          (-)      (-)                                         ______________________________________                                    

As is apparent from the test results shown in Table 2, it is shown that(R)-3,6-diamino-N-(2-aminoethyl) hexanamide (Ia) and(R)-3,6-diamino-N-(3-aminopropyl) hexanamide (Ib) according to thepresent invention have anti-HIV activities comparable to or sometimesuperior to those of D-β-lysylmethanediamine (Bellenamine) as acomparative drug, and they do not have cytotoxicity.

This indicates that the addition and co-existence of an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide according to this inventiondoes not allow HIV to proliferate in human MT-4 cells but can inhibitdevelopment of HIV antigen in the MT-4 cells. The novel compounds of theformula (I) according to this invention have therefore been confirmed toexhibit strong inhibitory activities agsint the infection of humanT-cells with HIV.

3! Acute Toxicity

In tests for estimation of acute toxicity by intravenous injection tomice, the novel compounds of the formula (I) according to thisinvention, particularly (R)-3,6-diamino-N-(2-aminoethyl)hexanamide (Ia)and (R)-3,6-diamino-N-(3-aminopropyl)hexanamide (Ib) did not cause deathof mice at a dosage of 250 mg/kg. As a result, these compounds aretherefore proved to have a low toxicity.

4! Antitumor Activity

The (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide of the general formula(I) according to this invention can further exhibit an antitumoractivity against the growth of tumor cells. To demonstrate this, mouseleukemia P388 cells as representative tumor cells were incubated in aculture medium in the presence of(R)-3,6-diamino-N-(2-aminoethyl)hexanamide (Ia) or(R)-3,6-diamino-N-(3-aminopropyl)hexanamide (Ib) as added at differentconcentrations of the tested compound. In this way, such concentrationof the tested compound which can inhibit the growth of the P388 cells by50%, that is, the IC₅₀ -value of the tested compound against the P388cells was estimated. For comparison purpose, Bellenamine was tested inthe same manner as above. The test results obtained are summarized inTable 3 below.

                  TABLE 3                                                         ______________________________________                                        Tested Compound  IC.sub.50 -value (μg/ml)                                  ______________________________________                                        Invention Compound (Ia)                                                                        0.012                                                        Invention Compound (Ib)                                                                        0.028                                                        Bellenamine (comparative)                                                                      0.36                                                         ______________________________________                                    

From the results of Table 3, it is shown that the tested compound ofthis invention can exhibit a higher antitumor activity than Bellenamineagainst the mouse leukemia P388 tumor cells.

Next, the production of the novel compound of the general formula (I)according to the present invention will be described in details.

In the process for the preparation of the novel compound of the generalformula (I) according to the aforesaid second aspect of this invention,there is employed as the starting compound such a mono-amino-protectedderivative of the α,ω-alkanediamine as represented by the generalformula (II) shown hereinbefore. This mono-amino-protected derivative ofthe formula (II) may be prepared by anyone of known various methods butpreferably may be prepared in accordance with the method of Atwell &Denny see the "Synthesis", page 1032 (1984)!. More specifically, forexample, an aqueous solution of propane-1,3-diamine is adjusted to pH3.8 with methane-sulfonic acid, followed by addition of ethanol. Theresulting solution is added dropwise with a solution ofbenzyloxycarbonyl chloride in dimethoxyethane under stirring at 20° C.At the same time, to the resulting mixture is added an aqueous solutionof potassium acetate so that the pH of the reaction mixture ismaintained at 3.5-4.5 during the reaction, whereby one of the aminogroups of the propane-1,3-diamine can be selectivelybenzyloxycarbonylated to affordN-(benzyloxycarbonyl)-propane-1,3-diamine in a high yield (seeReferential Example 2 given hereinafter).

The bis(N-aralkyloxycarbonyl)-D-β-lysine of the formula (III) to be usedas the reactant may be bis(N-benzyloxycarbonyl)-D-β-lysine (seeReferential Example 3 given hereinafter) which is available by themethod of the present inventors see "J. Antibiotics", 45, 1677(1992)!.

In the process of the second aspect of the present invention, thereaction for condensation of the compound of the formula (II) with thecompound of the formula (III) via the amido-bond can be conducted byusing a known method such as the active ester method, the acid halidemethod or the like.

As the amino-protecting groups for use in the process of the secondaspect of the invention, there can be employed various knownamino-protecting groups, including alkyloxycarbonyl groups. It ishowever desirable to select such amino-protecting groups which canreadily be removed under mild reaction conditions enough to avoid anyfurther change to occur in the reaction product as formed. Preferredexamples of the amino-protecting group include aralkyloxycarbonylgroups, such as benzyloxycarbonyl, para-methoxybenzyloxycarbonyl and thelike, which can be eliminated easily by hydrogenolysis.

An (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide of the general formula (I)according to the present invention or a pharmaceutically acceptable saltthereof may be admixed with a pharmaceutically acceptable solid orliquid carrier so as to form a pharmaceutical composition comprising thecompound of the formula (I) or the salt thereof as the activeingredient, which may be administered orally or parenterally.

According to a third aspect of the present invention, therefore, thereis provided a pharmaceutical composition, which comprises as activeingredient an (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide represented bythe general formula (I): ##STR10## wherein n stands for an integer of2-5, or an acid addition salt thereof, in association with apharmaceutically acceptable carrier for the active ingredient.

The pharmaceutical composition according to the third aspect of thepresent invention may be prepared in the form of various formulationssuch as injections, oral preparations, suppositories or the likecontaining the active compound of the formula (I) or a salt thereof asmixed with a pharmaceutically acceptable carrier or excipient. Anypharmaceutically acceptable carrier or excipient are selectable for thatpurpose. The nature and composition of the carrier used may varydepending on the administration route and manner. For example, water,ethanol, an animal or vegetable oil such as soybean oil, sesame oil ormineral oil, or a synthetic oil may be used as a liquid carrier. Usablesolid carriers include, for example, a sugar such as maltose or sucrose,an amino acid such as lysine, a cellulose derivative such ashydroxypropylcellulose, a polysaccharide such as cyclodextrin, a salt ofan organic acid such as magnesium stearate, or the like.

For the injections being prepared, it is generally preferable that theliquid medium of the injections comprises physiological saline, variousbuffered solutions, an aqueous solution of a sugar such as glucose,inositol or mannitol, or a glycol such as ethylene glycol orpolyethylene glycol. It is also feasible to formulate a lyophilizedpreparation containing the compound of the formula (I) or a salt thereofas the active ingredient in association with an excipient, e.g., a sugarsuch as inositol, mannitol, glucose, mannose, maltose or sucrose or anamino acid such as phenylalanine. Upon administration, such lyophilizedpreparation may be dissolved in a suitable solvent for injection, forexample, sterilized water or an intravenously-administrable liquid suchas physiological saline, aqueous solution of glucose, an aqueoussolution of electrolytes or an aqueous solution of amino acids.

Although the proportion of the compound of the formula (I) present inthe formulated composition may widely vary from one preparation toanother preparations, it may generally be in a range of 0.1-95 % byweight, preferably 1-90 % by weight. Upon the preparation of aninjection, for example, it is generally desirable that the injectionablesolution contains the compound of the formula (I) as active ingredientat a concentration of 0.1-5 % by weight. For oral administration, thecompound (I) as the active ingredient may be formulated into tablets,capsules, a powder, granules in combination with the solid carrier ormay be formulated into a solution, a dry syrup or the like incombination with the liquid carrier. In capsules, tablets, granules or apowder, the proportion of the compound of the formula (I) as the activeingredient present therein may usually be in a range of about 3-95 wt.%,preferably 5-90 wt.%, with the balance being formed of a carrier.

The dosage of the compound of the formula (I) may suitably be determinedin account of the age, body weight, symptom of patients and therapeuticpurpose as intended. The therapeutic, i.e., effective dosage of thecompound of the formula (I) may be generally in a range of 1-100mg/kg/day for the parenteral administration and in a range of 5-500mg/kg/day for the oral administration. This dosage can be administeredeither continuously or intermittently as long as the total dosage doesnot exceed such a spcific level that was decided in view of results ofanimal tests and various circumstances.

Similarly, the total dosage of the compound of the formula (I) given bythe parenteral administration may vary suitably depending on the way ofadministration, conditions of the patient or animal under treatment, forexample, the age, body weight, sex, sensitivity, foods or feed,administration time, administration route, drugs adminsteredconcurrently, conditions of the patient and disease. The suitable dosageand administration frequency of the compound of the formula (I) undergiven conditions must be determined by an expert physician through thetests of determining optimal dosage and in light of the aboveguidelines. These requirements for administration should also apply tothe oral administration of the compound of the formula (I).

In a further aspect, the present invention embraces use of an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide of the general formula (I) asdefined hereinbefore or a pharmaceutically acceptable salt thereof, inthe manufacture of a pharmaceutical composition for inhibiting infectionof human T-cells with AIDS virus.

In another aspect, the present invention includes use of an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide of the general formula (I) asdefined above or a pharmaceutically acceptable salt thereof, in themanufacture of an antibacterial composition.

In a still another aspect, the present invention includes use of an(R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamide of the general formula (I)as defined above or a pharmaceutically acceptable salt thereof, in themanufacture of an antitumor composition.

The present invention will hereinafter be illustrated with reference tothe following Referential Examples and Examples. It should however benoticed that this invention is in no way limited to or by the Examples.

REFERENTIAL EXAMPLE 1

Preparation of N-(benzyloxycarbonyl)ethylenediamine This preparation wasconducted in accordance with the method proposed by Atwell et al."Synthesis", pp. 1032-1033(1984)!

In 7 ml of water were dissolved 2.4 ml (35.9 mmol) of ethylenediamine.An aqueous solution (7 ml) of methanesulfonic acid (4.3 ml, 66.3 mmol)was added to the resulting solution to adjust its pH to 3.7. The aqueoussolution obtained was diluted with 20 ml of ethanol, followed bydropwise addition thereto of a solution (7 ml) of benzyl chloroformate(4.5 ml, 31.5 mmol) in dimethoxy-ethane under vigorous stirring at 20°C. At the same time, a 50%(w/v) aqueous solution of potassium acetatewas added dropwise to the resulting reaction mixture so that the pH ofthe reaction mixture was maintained at 3.5-4.5. After the addition ofbenzyl chloroformate, the resulting mixture was stirred for one hour atroom temperature. The reaction solution so obtained was thenconcentrated to dryness.

To the residue so obtained were added 70 ml of water. The resultingmixture was stirred and then, an insoluble matter was filtered offtherefrom. The filtrate was washed thrice with benzene (20 ml), adjustedto pH 10 or more with a 10N aqueous solution of sodium hydroxide andthen extracted twice with benzene (40 ml). The benzene layer (theextracts) obtained was washed with 40 ml of a saturated aqueous solutionof sodium chloride, dried over anhydrous sodium sulfate and concentratedto dryness, whereby 4.30 g of a crude product of the above-titledcompound were obtained. This crude product obtained was loaded on anupper part of a silica gel column (430 g, "C-300"; trade name, a productof Wako Purechemical Industries, Ltd.) which had been packed with a 5:1mixed solvent of chloroform and methanol in advance, and the column wasthen developed using the same mixed solvent as a developer. Fractions ofthe eluate which showed a ninhydrin-positive single spot on TLC werecombined together and concentrated to dryness. Thus, 3.90 g ofN-(benzyloxycarbonyl)ethylenediamine were afforded as a colorless paste.

REFERENTIAL EXAMPLE 2

Preparation of N-(benzyloxycarbonyl)propane-1,3-diamine

This preparation was conducted again in accordance with the methodproposed by Atwell et al. "Synthesis", pp. 1032-1033(1984)!

In 7 ml of water were dissolved 3.0 ml (35.9 mmol) ofpropane-1,3-diamine. An aqueous solution (7 ml) of methanesulfonic acid(4.3 ml, 66.3 mmol) was added to the resulting solution to adjust its pHto 3.8. The aqueous solution obtained was diluted with 19.5 ml ofethanol, followed by dropwise addition of a solution (7 ml) of benzylchloroformate (4.5 ml, 31.5 mmol) in dimethoxyethane under vigorousstirring at 20° C. At the same time, a 50%(w/v) aqueous solution ofpotassium acetate was added dropwise to the resulting reaction mixtureso that the pH of the reaction mixture was maintained at 3.5-4.5. Afterthe addition of benzyl chloroformate, the resulting mixture was stirredfor one hour at room temperature. The reaction solution so obtained wasthen concentrated to dryness.

To the residue obtained were added 70 ml of water. After the resultingmixture was stirred, an insoluble matter was filtered off therefrom. Thefiltrate was washed thrice with benzene (21 ml), adjusted to pH 10 ormore with a 10N aqueous solution of sodium hydroxide and then extractedtwice with benzene (40 ml). The benzene layer (the extracts) obtainedwas washed with 40 ml of a saturated aqueous solution of sodiumchloride, dried over anhydrous sodium sulfate and concentrated todryness, whereby 3.34 g of a crude product of the above titled compoundwere obtained. This crude product obtained was loaded on an upper partof a silica gel column (330 g, "C-300"; trade name, a product of WakoPurechemical Industries, Ltd.) which had been packed with a 5:1 mixedsolvent of chloroform and methanol in advance, and the column was thendeveloped using the same mixed solvent as a developer. Fractions of theeluate which showed a ninhydrin-positive single spot on TLC werecombined together and concentrated to dryness. Thus, 2.95 g ofN-(benzyloxycarbonyl)propane-1,3-diamine were afforded as a colorlesspaste.

REFERENTIAL EXAMPLE 3

Preparation of bis(N-benzyloxycarbonyl)-D-β-lysine

In 50 % aqueous methanol were dissolved 68.7 mg (0.47 mmol) ofD-β-lysine, and the resulting solution was added with 399.5 mg (1.46mmol) of benzyl S-4,6-dimethyl-pyrimid-2-ylthiocarbonate (product ofKokusan Chemical Works, Ltd.) and 0.135 ml (0.96 mmol) of triethylamine.The resulting mixture was stirred overnight at room temperature. Thereaction solution obtained was concentrated to dryness, followed bywashing the residue with aqueous methanol, to give 150mg of(N-benzyloxycarbonyl)-D-β-lysine as colorless powder (yield: 77%).

EXAMPLE 1

Preparation of(R)-3,6-bis(benzyloxycarbonylamino)-N-(2-benzyloxycarbonylaminoethyl)hexanamide(corresponding to the compound of the formula (IV) where A represents abenzyloxycarbonyl group and n stands for 2)

In 12 ml of dioxane were dissolved 401 mg (0.968 mmol) ofbis(N-benzyloxycarbonyl)-D-β-lysine, and the resultant solution was thenadded with 124 mg (1.077 mmol) of N-hydroxysuccinimide and 225 mg (1.090mmol) of dicyclo-hexylcarbodiimide. The resulting mixture was stirred atroom temperature for 21 hours, and the precipitate as formed was thenfiltered off therefrom. To the resulting filtrate, there were added asuspension (6 ml) of N-(benzyloxycarbonyl)ethylenediamine (284 mg, 1,460mmol) in dioxane and an aqueous solution (10 ml) of 125 mg (1.488 mmol)of sodium hydrogen carbonate, followed by stirring the resulting mixtureat room temperature for 3 hours.

The precipitate as produced was collected by filtration. The precipitatewas washed successively with 5 ml of water and 5 ml of cold dioxane andthen dried to yield 565 mg of a crude powder. The crude powder obtainedwas loaded on an upper part of a silica gel column (56 g, "C-300"; tradename, a product of Wako Purechemical Industries, Ltd.), which had beenpacked with a 20:1 mixed solvent of chloroform and methanol in advance,and the column was then subjected to development using the same mixedsolvent as a developer. Fractions of the eluate which showed aninhydrin-positive single spot on TLC were combined together andconcentrated to dryness. In this way, 530 mg of the titled compound wereobtained as a colorless powder.

    SI-MS: m/z 591 (MH.sup.+),  α!.sub.D.sup.24 +3.2°(c 1.08, DMSO)

EXAMPLE 2

Preparation of (R)-3,6-diamino-N-(2-aminoethyl)-hexanamide of theformula (Ia):

In 180 ml of methanol were suspended 1.41 g (2.39 mmol) of(R)-3,6-bis(benzyloxycarbonylamino)-N-(2-benzyloxycarbonylaminoethyl)hexanamide.To the resulting suspension was added a suspension of 718 mg of 10%palladium-carbon in 20 ml of water. After stirring, hydrogen gas waspassed through the resulting mixture at room temperature for 3 hours toeffect the reaction. The palladium-corbon was filtered off from thereaction mixture obtained, and the filtrate was concentrated to dryness.The residue obtained was dissolved in 15 ml of water and then subjectedto a chromatography on a column of "Amberlite CG-50" (trade name, NH₄ ⁺-form, 50 ml). After the column was washed with 200 ml of water, thecolumn was eluted with aqueous ammonia at concentrations gradientlyvarying from 0.5% to 4.0% (0.5%-increased steps; 200 ml at eachconcentration). Fractions of the eluate as eluted with 2.5% to 3.5%aqueous ammonia were combined together and concentrated to dryness, toafford 445 mg of (R)-3,6-diamino-N-(2-aminoethyl)hexanamide as acolorless paste.

EXAMPLE 3

Preparation of(R)-3,6-bis(benzyloxycarbonylamino)-N-(3-benzyloxycarbonylaminopropyl)hexanamide(corresponding to the compound of the formula (IV) where A represents abenzyloxycarbonyl group and n stands for 3)

In 4 ml of dioxane were dissolved 195 mg (0.926 mmol) ofbis(N-benzyloxycarbonyl)-D-β-lysine were dissolved, and the resultingsolution was added with 43.0 mg (0.374 mmol) of N-hydroxysuccinimide and74.0 g (0.359 mmol) of dicyclohexylcarbodiimide. The resulting mixturewas stirred at room temperature for 18 hours, and precipitate as formedwas filtered off therefrom. The resulting filtrate was added with asuspension (2 ml) of N-(benzyloxycarbonyl)propane-1,3-diamine (103 mg,0.495 mmol) in dioxane and an aqueous solution (3.5 ml) of sodiumhydrogen carbonate (44 mg, 0,524 mmol), followed by stirring theresultant mixture at room temperature for 4 hours. The reaction solutionso obtained was then added with 40 ml of water, followed by extractingtwice with 50 ml of chloroform. The chloroform layer (the extracts) wasdried over anhydrous sodium sulfate and then concentrated to dryness, togive 217 mg of a crude powder. This crude powder obtained was purifiedby preparative TLC ("Art. No. 5744"; trade name, product of Merck A.G.,Germany) with using a 20:1 mixed solvent of chloroform and methanol asthe eluent. Thus, 183 mg of the titled compound were obtained ascolorless powder.

    SI-MS: m/z 605(MH.sup.+),  α!.sub.D.sup.24 +1.3° (c 1.06, DMSO)

EXAMPLE 4

Preparation of (R)-3,6-diamino-N-(3-aminopropyl)-hexanamide of theformula (Ib)

In 18 ml of methanol were suspended 144 mg (0.238 mmol) of(R)-3,6-bis(benzyloxycarbonylamino)-N-(3-benzyloxycarbonylaminopropyl)hexanamide.To the resulting suspension was added a suspension of 81.0 mg of 10%palladium-carbon in 2 ml of water, followed by stirring. Hydrogen gaswas passed through the resulting mixture at room temperature for 3 hoursto effect the hydrogenolysis reaction. The palladium-carbon was filteredoff from the reaction mixture obtained, and the filtrate was thenconcentrated to dryness. The residue obtained was dissolved in 1 ml ofwater and the resulting solution was subjected to chromatography on acolumn of "Amberlite CG-50" (trade name, NH₄ ⁺ -form, 5 ml). After thecolumn was washed with 25 ml of water, the column was eluted withaqueous ammonia at concentrations gradiently varying from 0.5% to 7.0%(0.5%-increased steps; 25 ml at each concentration). Fractions of theeluate as eluted with 4.0% to 6.0% aqueous ammonia were combinedtogether and concentrated to dryness, to afford 48.2 mg of(R)-3,6-diamino-N-(3-aminopropyl)hexanamide as a colorless paste.

INDUSTRIAL APPLICAPABILITY OF INVENTION

The new compounds, (R)-3,6-diamino-N-(ω-aminoalkyl) hexanamide of thegeneral formula (I) are now synthesized in accordance with the presentinvention, as described hereinbefore. These new compounds and their acidaddition salts are chemically stable and have an activity of inhibitingthe growth of Gram-positive bacteria and Gram-negative bacteria and alsoan activity of inhibiting the infection of human T-cells with AIDSvirus, as well as an activity of inhibiting the growth of tumor cells,so that these new compounds are useful as a chemotherapeutic agent fortreatment of bacterial infections and HIV infection, and also as anantitumor agent.

We claim:
 1. An (R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide representedby the general formula (I): ##STR11## wherein n stands for an integer of2 or 3 and (R) means (R)-configuration, or an acid addition saltthereof.
 2. A compound according to claim 1 wherein n stands for 2 inthe general formula (I), which is(R)-3,6-diamino-N-(2-aminoethyl)hexanamide, or an acid addition saltthereof.
 3. A compound according to claim 1 wherein n stands for 3 inthe general formula (I), which is(R)-3,6-diamino-N-(3-aminopropyl)hexanamide, or an acid addition salthereof.
 4. A process for the preparation of an(R)-3,6-diamino-N-(ω-aminoalkyl)hexanamide represented by the generalformula (I): ##STR12## wherein n stands for an integer of 2 or 3 and (R)means (R)-configuration, which comprises the steps of condensing,through an amido-bond, a mono-amino-protected derivative of anα,ω-alkanediamine represented by the general formula (II):

    H.sub.2 N(CH.sub.2).sub.n NH-A                             (II)

wherein A represents an aralkyloxycarbonyl group which is anamino-protecting group easily removable by hydrogenolysis and n standsfor an integer of 2 or 3, with a bis(N-aralkyloxycarbonyl)-D-β-lysinerepresented by the general formula (III): ##STR13## wherein A has thesame meaning as described above, to form an(R)-3,6-bis(aralkyloxycarbonylamino)-N-(ω-aralkyloxycarbonylaminoalkyl)hexanamiderepresented by the following general formula (IV): ##STR14## wherein Aand n have the same meanings as described above, and then subjecting thecompound of the formula (IV) to hydrogenolysis to remove theamino-protecting aralkyloxycarbonyl groups therefrom.
 5. Apharmaceutical composition, which comprises as active ingredient an (R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamide represented by the generalformula (I): ##STR15## wherein n stands for an integer of 2 or 3 and (R)means (R)-configuration, or an acid addition salt thereof, inassociation with a pharmaceutically acceptable carrier for the activeingredient.
 6. The method of treating infection of a subject with AIDSvirus comprising: treating said subject with an(R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamide of the general formula (I)as defined in claim 1 or a pharmaceutically acceptable salt thereof. 7.The method of inhibiting the growth of bacteria comprising, treatingsaid bacteria with an (R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamide of thegeneral formula (I) as defined in claim 1 or a pharmaceuticallyacceptable salt thereof.
 8. The method of treating tumor growthcomprising, treating said tumor with an(R)-3,6-diamino-N-(ω-aminoalkyl)-hexanamide of the general formula (I)as defined in claim 1 or a pharmaceutically acceptable salt thereof.